Cyclic nucleotide gated Ca channels and non-self perception in plant pathogen defense responses

有机体适应性的一个基本方面涉及检测细胞外环境中“非自我”的存在。在病原微生物入侵的情况下，非自我存在的感知需要通过细胞信号转导级联进行翻译，以便受到攻击的生物体的细胞可以发起防御或免疫反应。该项目的目的是为植物细胞中病原体感知的早期事件提供新的见解。先前的研究表明，钙（Ca）通过环核苷酸门控阳离子通道（CNGC）蛋白流入细胞的细胞质，下游一氧化氮（NO）的产生对植物免疫应答信号级联反应至关重要。本项目的研究重点将利用我们对CNGC功能和调控相关分子结构的理解来阐明这一信号级联的进一步步骤。假设将被测试关于该CNGC从内向Ca传导上游的信号步骤；即将病原体感知与通道激活联系起来。工作将确定环核苷酸升高是否以及如何发生在植物细胞响应病原体识别。该项目的一个组成部分旨在检测一些植物细胞膜受体蛋白产生环核苷酸的可能能力，环核苷酸是通过CNGCs激活Ca传导的信号分子。其他研究将集中于鉴定可能影响CNGC通道激活的调节分子，从而调节植物对病原体的免疫反应。更广泛的影响。项目活动包括利用创新的分子遗传学教师培训方案进行外联和教育工作。这个为期两周的全天课程以综合的方式培训分子遗传学实验方法和基本原理的参与者。参与者经过培训，并提供了一套工具和资源（信息和生物），使他们能够在高中生物课上展示分子生物学体验实验室教育计划。该计划包括一个包（开发使用作为本项目的一部分）的PowerPoint演示文稿，蛋白质建模软件，实验课程计划，教师指南，80页的分子遗传学实验室手册，一个注释目录的网络资源，以帮助高中教师获取生物和信息资源，等等。作为该项目拟议的更广泛影响的一部分，基于这些材料的为期两周的培训计划将在资助期间每年夏天向教育学院的五名本科生和五名现有的高中生物教师提供。当高中教师在全州的学校实施该计划时，将向他们提供援助。教师培训计划和高中实验课程的所有信息都将发布在一个URL上，并通过这个网站和其他网站向广大高中生物教师提供。该项目的教育部分包括继续培训一名当前的非裔美国女性研究生，她将承担该项目的研究目标，作为其博士学位的一部分。

A fundamental aspect of organism fitness involves the detection of a "non-self" presence in the extracellular environment. In the case of a pathogenic microbe invader, perception of non-self presence needs to be translated through a cellular signal transduction cascade so that the cells of the organism under attack can mount a defense or immune response. The aim of this project is to provide new insights into the early events of pathogen perception in plant cells. Prior research has shown that calcium (Ca) flow into the cytoplasm of cells through a cyclic nucleotide gated cation channel (CNGC) protein, and downstream nitric oxide (NO) generation were critical to the plant immune response signaling cascades. The research focus of this project will take advantage of our understanding of the molecular architecture related to CNGC function and regulation to elucidate further steps in this signaling cascade. Hypotheses will be tested regarding the signaling steps upstream from inward Ca conduction by this CNGC; i.e. linking pathogen perception to channel activation. Work will determine if and how cyclic nucleotide elevation occurs in plant cells responding to pathogen recognition. One component of the project aims to examine some plant cell membrane receptor proteins for possible ability to generate cyclic nucleotide, the signaling molecule that activates Ca conduction through CNGCs. Other studies will focus on identifying regulatory molecules that may impact activation of the CNGC channels, and therefore modulate plant immune response to pathogens. Broader impacts. Project activity includes use of an innovative molecular genetics teacher-training program for outreach and education efforts. This two-week full day program trains participants in molecular genetics experimental methods and underlying principles in an integrated fashion. Participants are trained, and provided with a package of tools and resources (informational and biological) to allow them to present a molecular biology experiential lab educational program in high school biology classes. This program includes a package (developed for use as part of this project) of PowerPoint presentations, protein modeling software, lab lesson plans, teacher's guides, an 80-page molecular genetics lab manual, an annotated catalog of web resources to assist high school teachers in obtaining biological and informational resources, and more. As part of the proposed broader impacts of the project, the two-week training program based on this material will be delivered to five undergraduate School of Education students as well as five current high school biology teachers each summer during the funding period. Assistance will be provided to high school teachers as they implement the program in schools throughout the State. All of the information in the teacher-training program and high school lab curriculum will be posted on a URL and made available to a wide audience of high school biology teachers through this and other web sites. The educational component of the project includes continued training of a current female African-American graduate student who will undertake research objectives of the project as part of her Ph.D. degree.